Dataset Information

Substrate selectivity of the sublancin S-glycosyltransferase.

ABSTRACT: SunS is a novel S-glycosyltransferase involved in the biosynthesis of the antimicrobial peptide sublancin. It selectively modifies Cys22 in a 56 amino acid peptide substrate SunA and can accept a variety of NDP sugars. This study reports the substrate selectivity with regard to the peptide substrate and the antimicrobial activity of the resulting sublancin analogues. The results suggest that SunS recognizes an ?-helix N-terminal of the Cys to be glycosylated, which is present in a flexible linker. Interestingly, when Cys22 is mutated, sugar attachment is not required for sublancin antimicrobial activity. Furthermore, the sublancin-producing strain Bacillus subtilis 168 also becomes susceptible to such mutants. These data suggest that S-glycosylation may be important for self-resistance.

SUBMITTER: Wang H

PROVIDER: S-EPMC3191765 | biostudies-literature | 2011 Oct

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Substrate selectivity of the sublancin S-glycosyltransferase.

Wang Huan H van der Donk Wilfred A WA

Journal of the American Chemical Society 20110921 41

SunS is a novel S-glycosyltransferase involved in the biosynthesis of the antimicrobial peptide sublancin. It selectively modifies Cys22 in a 56 amino acid peptide substrate SunA and can accept a variety of NDP sugars. This study reports the substrate selectivity with regard to the peptide substrate and the antimicrobial activity of the resulting sublancin analogues. The results suggest that SunS recognizes an α-helix N-terminal of the Cys to be glycosylated, which is present in a flexible linke ...[more]

PMID: 21910430

Similar Datasets

Project description:The polypeptide N-acetylgalactosaminyl transferase (GalNAc-T) enzyme family initiates O-linked mucin-type glycosylation. The family constitutes 20 isoenzymes in humans. GalNAc-Ts exhibit both redundancy and finely tuned specificity for a wide range of peptide substrates. In this work, we deciphered the sequence and structural motifs that determine the peptide substrate preferences for the GalNAc-T2 isoform. Our approach involved sampling and characterization of peptide-enzyme conformations obtained from Rosetta Monte Carlo-minimization-based flexible docking. We computationally scanned 19 amino acid residues at positions -1 and +1 of an eight-residue peptide substrate, which comprised a dataset of 361 (19x19) peptides with previously characterized experimental GalNAc-T2 glycosylation efficiencies. The calculations recapitulated experimental specificity data, successfully discriminating between glycosylatable and non-glycosylatable peptides with a probability of 96.5% (ROC-AUC score), a balanced accuracy of 85.5% and a false positive rate of 7.3%. The glycosylatable peptide substrates viz. peptides with proline, serine, threonine, and alanine at the -1 position of the peptide preferentially exhibited cognate sequon-like conformations. The preference for specific residues at the -1 position of the peptide was regulated by enzyme residues R362, K363, Q364, H365 and W331, which modulate the pocket size and specific enzyme-peptide interactions. For the +1 position of the peptide, enzyme residues K281 and K363 formed gating interactions with aromatics and glutamines at the +1 position of the peptide, leading to modes of peptide-binding sub-optimal for catalysis. Overall, our work revealed enzyme features that lead to the finely tuned specificity observed for a broad range of peptide substrates for the GalNAc-T2 enzyme. We anticipate that the key sequence and structural motifs can be extended to analyze specificities of other isoforms of the GalNAc-T family and can be used to guide design of variants with tailored specificity.

Dataset Information

Substrate selectivity of the sublancin S-glycosyltransferase.

Publications

Substrate selectivity of the sublancin S-glycosyltransferase.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets